Use of 4-H-1-benzopryan-4-one derivatives as inhibitors of smooth muscle cell proliferation

Abstract

The use of 4-H-1-benzopyran-4-one derivatives as inhibitors of smooth muscle cell proliferation. Smooth muscle cell (SMC) proliferation is a critical component of neointimal formation in many animal models of vascular injury, and in many human lesions as well. Cell cycle inhibition by gene transfer techniques can block SMC proliferation and lesion formation in many animal models, although these methods are not yet applicable to the treatment of human disease. Flavopiridol is a recently identified, potent, orally available cyclin-dependent kinase inhibitor. Given the role of smooth muscle cell (SMC) proliferation in vascular disease, we tested the effects of flavopiridol, a recently identified cyclin-dependent kinase inhibitor, on SMC growth in vitro and in vivo. Flavopiridol (75 nmol/L) potently blocked SMC proliferation, an effect that was associated with downregulation of cyclin-dependent kinase activity and cell cycle-related gene expression. We examined the effects of flavopiridol on SMC proliferation in vivo in the rat carotid injury model. Flavopiridol (5 mg/kg) decreased neointimal size by 35% and 39% at 7 and 14 days, respectively, after balloon injury. Flavopiridol may be a potential therapeutic tool in the treatment of SMC-rich vascular lesions. 4-H-1-benzopyran-4-one derivatives inhibit smooth muscle cell proliferation at low dosage levels.

What is claimed is:  
     
       1. A method of inhibiting smooth muscle cell proliferation comprising administering to a patient an effective amount of a compound of the formula I, which is less than 70% if the dosage to control tumor growth,                    
       wherein 
       R 1  is hydrogen, alkyl having 1 to 6 carbon atoms, aryl-C 1 -C 4 -alkyl; C 1 -C 6 -alkyl substituted by halogen, hydroxy, or carboxy; C 3 -C6-cycloalkyl, pyridyl, thienyl, C 3 -C 6 -cycloalkyl-C 1 -C 4 -alkyl, C 2 -C 6 -alkenyl, C 2 -C 6 -alkynyl, phenyl; phenyl, mono- or polysubstituted by halogen, C 1 -C 4 -alkyl, C 1 -C 4 -alkoxy, hydroxyl, carboxyl, COO-alkyl, CONH 2 , CONH-alkyl, CON(alkyl) 2 , nitro, trifluoromethyl, amino, C 1 -C 4 -alkylamino, di-C 1 -C 4 -alkylamino, or phenyl; naphthyl, carboxyl, —CHO, COO-C 1 -C 4 -alkyl, a primary amino, alkylamino, aralkylamino, dialkylamino, amido, arylamino, diarylamino, or —CH 2 O-C 1 -C 4 -alkyl;  
       R 2  is hydrogen, alkyl having 1 to 6 carbon atoms, aryl, nitro, amino, di-C 1 -C 4 -alkylamino, a halogen, hydroxyl, alkoxy, —COOH, —COO-C 1 -C 4 -alkyl, —CHO, —CH 2 OH or —CH 2 O-C 1 -C 4 -alkyl;  
       R 3  is hydrogen, C 1 -C 4 -alkyl; C 1 -C 4 -alkyl substituted by halogen, hydroxy or carboxy; hydroxyl, carboxyl, nitro, amino, C 1 -C 4 -alkylamino, di-C 1 -C 4 -alkylamino, halogen, —O-alkyl-C(O)-alkyl, —CHO, —CH 2 OH, —CH 2 O-C 1 -C 4 -alkyl or R 2 N—C(O)—O—, wherein R is H, C 1 -C 6 -alkyl, cycloalkyl; or —O-alkyl-C(O)-alkyl or aryl;  
       R 4  is hydrogen, hydroxyl C 1 -C 4 -alkoxy, C 1 -C 4 -alkanoyloxy, C 1 -C 4 -alkoxycarbonyl, aryloxy, amino, C 1 -C 4 -alkylamino, di-C 1 -C 4 -alkylamino, or  
       R′ 2 —N—C(O)—O—  
       wherein R′ is H, C 1 -C 6 -alkyl, cycloalkyl or aryl;  
       R 5  is hydrogen, C 1 -C 6 -alkyl, aryl-C 1 -C 4  alkyl, C 3 -C 6 -cycloalkyl, C 3 -C 6 -cycloalkyl-C 1 -C 4 -alkyl, alkylamino, C 1 -C 4 -alkanoyl, —C(O)—O-C 1 -C 4 -alkyl or aroyl, where the aryl group in R 1 , R 2 , R 3 , R 4 , and R 5  is unsubstituted phenyl or phenyl that is mono or polysubstituted by halogen, C 1 -C 4 -alkyl, C 1 -C 4 -alkoxy, hydroxyl, carboxyl, COO-alkyl, CONH 2 , CONH-alkyl, CON(alkyl) 2 , nitro, trifluoromethyl, amino, C 1 -C 4 -alkylamino, di-C 1 -C 4 -alkylamino or phenyl; m is an integer between 0 and 3 and n is 1,  
       or a pharmacologically acceptable acid addition salt thereof.  
     
     
       2. The method of  claim 1 , wherein the effective amount of a compound of the formula I is less than 60% of the dosage to control tumor growth. 
     
     
       3. The method of  claim 2 , wherein the effective amount of a compound of the formula I is less than 50% of the dosage to control tumor growth. 
     
     
       4. The method of  claim 1 , wherein the compound is a compound of formula Ia compounds of the formula Ia, is less than which is less than 70% of the dosage to control tumor growth,                    
       in which R 1  is hydrogen, C 1 -C 3 -alkyl, naphthyl, phenyl; phenyl mono- or polysubstituted by halogen, C 1 -C 4 -alkyl, C 1 -C 4 -alkoxy, hydroxyl, carboxyl, COO-alkyl, CONH 2 , CONH-alkyl, CON(alkyl) 2 , nitro, trifluoromethyl, amino, C 1 -C 4 -alkylamino, di-C 1 -C 4 -alkylamino, or phenyl; pyridyl, or thienyl; 
       R 2  is hydrogen or C 1 -C 3 -alkyl;  
       R 5  is C 1 -C 3 -alkyl, C 3 -C 5 -cycloalkyl, or C 3 -C 5 -cycloalkyl-C 1 -C 4 -alkyl.  
     
     
       5. The method of  claim 4 , wherein the substituents of formula la have the following meanings 
       R 1  is phenyl, thienyl, pyridyl, chlorophenyl, dichlorophenyl, methylphenyl, aminophenyl, bromophenyl, hydroxyphenyl or naphthyl;  
       R 2  is hydrogen and  
       R 5  is methyl.  
     
     
       6. The method of  claim 5 , in which the compound is (−)-cis,-5,7-dihydroxy-2-) (2-chlorophenyl)-8-[4-(3-hydroxy-1-methyl)-piperidinyl]-4H-benzopyran-4-one (Flavopiridol). 
     
     
       7. The method of  claim 4 , wherein the effective amount of a compound of the formula Ia is less than 60% of the dosage to control tumor growth. 
     
     
       8. The method of  claim 7 , wherein the substituents of formula Ia have the following meanings 
       R 1  is phenyl, thienyl, pyridyl, cholorophenyl, dichlorophenyl, methylphenyl, aminophenyl, bromophenyl, hydroxyphenyl or naphthyl;  
       R 2  is hydrogen and  
       R 5  is methyl.  
     
     
       9. The method of  claim 8 , in which the compound is (−)-cis,-5,7-dihydroxy-2-) (2-chlorophenyl)-8-[4-(3-hydroxy-1-methyl)-piperidinyl]-4H-benzopyran-4-one (Flavopiridol). 
     
     
       10. The method of  claim 8 , wherein the effective amount of a compound of the formula Ia is less than 50% of the dosage to control tumor growth. 
     
     
       11. The method of  claim 11 , wherein the substituents of formula Ia have the following meanings 
       R 1  is phenyl, thienyl, pyridyl, chlorophenyl, dichlorophenyl, methylphenyl, aminophenyl, bromophenyl, hydroxyphenyl or naphthyl;  
       R 2  is hydrogen and  
       R 5  is methyl.  
     
     
       12. The method of  claim 11 , in which the compound is (−)-cis,-5,7-dihydroxy-2-) (2-chlorophenyl)-8-[4-(3-hydroxy-1-methyl)-piperidinyl]-4H-benzopyran-4-one (Flavopiridol). 
     
     
       13. The method of  claims 1  to  12 , wherein the administration is for treatment of smooth muscle cell-rich vascular lesions. 
     
     
       14. The method of  claims 1  to  12 , wherein the administration is for treatment of lesions after balloon injury. 
     
     
       15. The method of  claims 1  to  12 , wherein the administration is for treatment after stent implantation.